实时荧光定量PCR检测结直肠腺癌患者组织中广泛型线粒体肌酸激酶基因的表达

Real-time Fluorescence Quantitative PCR in Detecting uMtCk Gene Expression in Colorectal Adeno Carcinoma

目的建立检测人广泛型线粒体肌酸激酶（ubiquitous mitochondrial creatine kinase，uMtCk）mRNA荧光定量的方法，并检测了结直肠腺癌病人肿瘤组织和癌旁组织中uMtCk基因表达的水平。方法基于TaqMan荧光探针技术，以GAPDH作为内参，建立实时荧光定量RT-PCR方法，并检测了30名结直肠腺癌病人肿瘤组织和癌旁组织uMtCk mRNA的表达。结果30倒结直肠肿瘤患者肿瘤组织中有uMtCk mRNA的表达，范围为7.4×10^5－5.3×10^8拷见／μg RNA，均值为（9.4±5.7）×10^6拷贝／μg RNA；30例癌旁组织uMtCk表达为4.5×10^3－6.9×10^5拷贝／μg RNA，均值为（8.6±3.4）×10^4拷贝／μg RNA。结论成功地建立了人uMtCk基因表达含量的荧光定量检测方法，检测结果可用绝对拷贝数表示，定量准确可靠。且结直肠腺癌病人肿瘤组织uMtCk mRNA的含量是升高的，提示uMtCk mRNA含量的检测有助于结直肠腺癌病人临床诊断。

Objective To establish a specific fluorogenic quantitative RT-PCR （FQ-RT-PCR） method for detecting the expression of uMtCk（ubiquitous mitochondrial creatine kinase）gene in both tumor tissues and non-carcinoma adjacent tissues of colorectal adeno carcinoma and investigate the relationship between uMtCk mRNA expression and colorectal adeno carcinoma. Methods Based on fluorescent TaqMan methodology, GAPDN as endogenous control, a real-time quantitative RT-PCR was set up. The uMtCk gene expression levels in both carcinoma tissues and non-carclnoma adjacent tissues of 30 patients with colorectal adeno carcinoma were measured by real-time RT-PCR. Results In 30 colorectal adeno carcinoma patlents,the uMtCk mRNA expression ranged from 7.4×10^5-5.3×10^8 copy/μg RNA,the mean value was（9.4±5.7）×10^6 copy/μg RNA. In 30 non-carcinoma adjacent tissues the uMtCk mRNA expression ranged from 4.5×10^3-6.9×10^5 copy/μg RNA,the mean level was （8.6±3.4）×10^4 copy/μg RNA. Conclusion uMtCk gene expression detected with FQ-RT-PCR had an accurate,speedy results. The uMtCk mRNA expression level was significantly elevated in carcinoma tissues of colorectal adeno carcinoma. The uMtCk mRNA might be useful for diagnosis of colorectal adeno carcinoma.